LOCUS       sequence-334368-       10415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded lentiviral vector to express CDK4 in mammalian cells under 
            control of EF1a promoter. Barcoded for transcript detection in 
            single cell RNA-seq.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10415)
  AUTHORS   Parekh U, McDonald D, Dailamy A, Wu Y, Cordes T, Zhang K, Tipps A, 
            Metallo C, Mali P
  TITLE     Charting oncogenicity of genes and variants across lineages via 
            multiplexed screens in teratomas.
  JOURNAL   iScience. 2021 Sep 20;24(10):103149. doi: 
            10.1016/j.isci.2021.103149. eCollection 2021 Oct 22.
  PUBMED    34646987
REFERENCE   2  (bases 1 to 10415)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "iScience.";
            date: "2021-09-20"; volume: "24(10):103149. doi"; pages: " 
            10.1016/j.isci.2021.103149. eCollection 2021 Oct 22"
COMMENT     Sequence Label: LV_EF1a_CDK4-P2A-Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..10415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2693..3871
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2924..3862
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3819..3839
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3888..3907
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3888..3906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3964..3973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4888..4944
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4945..5970
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4945..5970
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(6162..6195)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             6220..6400
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6426..6443)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6432..6656
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6702..7130
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6789..6808)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6999..7020
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7139..7159)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7144..7473
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7324..7459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7386..7405
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7521..7568
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7587..7961
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8091..8212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8128..8147)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8182..8201
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8261..8277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8261..8277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8274..8296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8285..8301
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8309..8315,8316..8333,8334..8339))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8354..8375
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8492..8509)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8663..9251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8743..8762)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9422..10213,10214..10282))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10045..10064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10283..10387)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atcgataagc tttgcaaaga tggataaagt tttaaacaga
     2641 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
     2701 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     2761 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     2821 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     2881 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     2941 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     3001 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     3061 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     3121 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     3181 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     3241 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     3301 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     3361 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     3421 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     3481 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     3541 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     3601 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     3661 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     3721 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     3781 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     3841 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     3901 tatagggaga cccaagctgg ctaggtaagc ttgatatcga attcctgcag cccgggggat
     3961 cccgccacca tggctacctc tcgatatgag ccagtggctg aaattggtgt cggtgcctat
     4021 gggacagtgt acaaggcccg tgatccccac agtggccact ttgtggccct caagagtgtg
     4081 agagtcccca atggaggagg aggtggagga ggccttccca tcagcacagt tcgtgaggtg
     4141 gctttactga ggcgactgga ggcttttgag catcccaatg ttgtccggct gatggacgtc
     4201 tgtgccacat cccgaactga ccgggagatc aaggtaaccc tggtgtttga gcatgtagac
     4261 caggacctaa ggacatatct ggacaaggca cccccaccag gcttgccagc cgaaacgatc
     4321 aaggatctga tgcgccagtt tctaagaggc ctagatttcc ttcatgccaa ttgcatcgtt
     4381 caccgagatc tgaagccaga gaacattctg gtgacaagtg gtggaacagt caagctggct
     4441 gactttggcc tggccagaat ctacagctac cagatggcac ttacacccgt ggttgttaca
     4501 ctctggtacc gagctcccga agttcttctg cagtccacat atgcaacacc tgtggacatg
     4561 tggagtgttg gctgtatctt tgcagagatg tttcgtcgaa agcctctctt ctgtggaaac
     4621 tctgaagccg accagttggg caaaatcttt gacctgattg ggctgcctcc agaggatgac
     4681 tggcctcgag atgtatccct gccccgtgga gcctttcccc ccagagggcc ccgcccagtg
     4741 cagtcggtgg tacctgagat ggaggagtcg ggagcacagc tgctgctgga aatgctgact
     4801 tttaacccac acaagcgaat ctctgccttt cgagctctgc agcactctta tctacataag
     4861 gatgaaggta atccggaggg atccggcgca acaaacttct ctctgctgaa acaagccgga
     4921 gatgtcgaag agaatcctgg accgatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     4981 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     5041 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     5101 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     5161 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     5221 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     5281 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     5341 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     5401 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     5461 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     5521 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     5581 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     5641 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     5701 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     5761 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     5821 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     5881 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     5941 cccagcactc gtccgagggc aaaggaataa gttagaacta tttcctggct gttacgcgga
     6001 cgcctagtgt acagcgcact actgacgggc ttgttaaaca gtgaaccgcc cgccccacga
     6061 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgaacaaaa gaaaaggggg
     6121 gactggaagg gctaattcac tcccaacgaa gacaagatat cataacttcg tatagcatac
     6181 attatacgaa gttatcggct agctggtccg gactgtactg ggtctctctg gttagaccag
     6241 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     6301 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     6361 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
     6421 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     6481 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6541 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6601 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     6661 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     6721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     7021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     7081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     7141 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     7201 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     7261 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     7321 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     7381 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     7441 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     7501 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     7561 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     7621 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     7681 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     7741 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     7801 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     7861 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     7921 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     7981 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     8041 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     8101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8221 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     8281 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     8341 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     8401 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     8461 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     8521 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8581 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8641 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8701 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8761 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8821 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8881 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8941 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     9001 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     9061 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     9121 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     9181 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     9241 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     9301 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     9361 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    10021 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    10081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    10141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    10201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    10261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
    10321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    10381 ttccgcgcac atttccccga aaagtgccac ctgac
//
